Murine retroviruses re-engineered for lineage tracing and expression of toxic genes in the developing chick embryo.
We describe two replication incompetent retroviral vectors that co-express green fluorescent protein (GFP) and beta-galactosidase. These vectors incorporate either the avian reticuloendotheliosis (spleen necrosis virus; SNV) promoter or the chick beta-actin promoter, into the backbone of the murine leukemia (MLV) viral vector. The additional promoters drive transgene expression in avian tissue. The remainder of the vector is MLV-like, allowing high titer viral particle production by means of transient transfection. The SNV promoter produces high and early expression of introduced genes, enabling detection of the single copy integrated GFP gene in infected cells and their progeny in vivo. Substitution of the LacZ coding DNA with a relevant gene of interest will enable its co-expression with GFP, thus allowing visualization of the effect of specific and stable changes in gene expression throughout development. As the VSV-G pseudotyped viral vector is replication incompetent, changes in gene expression can be controlled temporally, by altering the timing of introduction.
Venters
Sara J
SJ
Department of Neurosurgery, University of California, San Francisco, California 94143, USA.
Dias da Silva
Magnus R
MR
Hyer
Jeanette
J
D000199:Actins / Q000235:genetics; D000818:Animals; D002642:Chick Embryo; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics*; D051379:Mice; D008979:Moloney murine leukemia virus / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D053079:Trager duck spleen necrosis virus / Q000235:genetics; D019076:Transgenes / Q000235:genetics*; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics*
D000199:Actins; D049452:Green Fluorescent Proteins; D001616:beta-Galactosidase